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Journal of Stress Physiology & Biochemistry, Vol. 10 No. 1 2014, pp. 14-25 ISSN 1997-0838Original Text Copyright © 2014 by Aloui, Souguir, Hannachi
ORIGINAL ARTICLE
Effect of Salt Stress (NaCl) on Germination and Early Seedling
Parameters of Three Pepper Cultivars (Capsicum annuum L.)
Aloui Hassen 1, Souguir Maher 2*, and Hannachi Cherif 2
1 Faculty of Sciences of Bizerte, University of Carthage 7021 Zarzouna, Tunisia2 Department of Horticultural sciences, High Institute of Agronomy, Chott-Mariem, 4042 Sousse,
Tunisia
*E-Mail: mahermaster11@yahoo.fr
Received August 4, 2013
Salinity is one of the major environmental problem that lead to a deterioration of agricultural land and, as a result, to a reduction in crop productivity worldwide. This research tested the effect of different salinity levels on germination and early seedling growth of three pepper (Capsicum annuum L.) cultivars which were "Beldi", "Baklouti" and "Anaheim Chili". Experimental treatment included 7 concentrations of NaCl (0, 2, 4, 6, 8, 10 and 12 g/l). Results indicated that all investigate traits were affected by salt stress. Salt stress affected on germination parameters and radicle and plumule length. Fresh weight and dry weight of evaluated seedlings was also affected. "Anaheim Chili" cultivar was shown to be the most restraint cultivar to salt stress in comparison to "Beldi" and "Baklouti" cultivars.
Key words: pepper cultivars , germination parameters, salt stress
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
Effect of Salt Stress (NaCl) on Germination...
ORIGINAL ARTICLE
Effect of Salt Stress (NaCl) on Germination and Early Seedling
Parameters of Three Pepper Cultivars (Capsicum annuum L.)
Aloui Hassen 1, Souguir Maher 2*, and Hannachi Cherif 2
1 Faculty of Sciences of Bizerte, University of Carthage 7021 Zarzouna, Tunisia2 Department of Horticultural sciences, High Institute of Agronomy, Chott-Mariem, 4042 Sousse,
Tunisia
*E-Mail: mahermaster11@yahoo.fr
Received August 4, 2013
Salinity is one of the major environmental problem that lead to a deterioration of agricultural land and, as a result, to a reduction in crop productivity worldwide. This research tested the effect of different salinity levels on germination and early seedling growth of three pepper (Capsicum annuum L.) cultivars which were "Beldi", "Baklouti" and "Anaheim Chili". Experimental treatment included 7 concentrations of NaCl (0, 2, 4, 6, 8, 10 and 12 g/l). Results indicated that all investigate traits were affected by salt stress. Salt stress affected on germination parameters and radicle and plumule length. Fresh weight and dry weight of evaluated seedlings was also affected. "Anaheim Chili" cultivar was shown to be the most restraint cultivar to salt stress in comparison to "Beldi" and "Baklouti" cultivars.
Key words: pepper cultivars , germination parameters, salt stress
Salt stress is certainly one of the most serious
environmental factors limiting the productivity of
crop plant (Ashraf, 1999). It is one of the major
abiotic stresses, and approximately 800 million
hectares of land are affected by high salt
concentrations throughout the world (Munns,
2005). Both osmotic and ionic stresses can cause
stunted growth and a reduced plant yield (Munns,
2002) and the irrigation with saline water is one of
the principal factors that lead to salt accumulation
and leads to a decrease in agriculture productivity.
Germination of seeds is a complex process
depending on genetic and environmental factors,
such as temperature, light, and salinity (Barbour,
1988; Mahmoud, 1985). In fact, salinity is one of
the most important factors limiting plant growth
and delaying seed germination as well as final
germination percentage (Rahman et al., 2000).
Salinity can affect germination and seedling growth
either by creating an osmotic pressure that
prevents water uptake or by the toxic effects of
sodium and chloride ions (Hopper et al., 1979).
Pepper (Capsicum annuum L.) is a glycophyte
plant which can't survive under high salinity stress
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
15
Aloui et al.
or can survive with affected yields (Ibn Maaouia-
Houimli et al., 2008) . Certainly , salinity reduces
performances especially in arid and semi-arid
regions characterized by irregular and insufficient
rainfall (Munns et al., 2006). In Tunisia, pepper
(Capsicum annuum L.) is an important crop
considered as the fifth vegetable speculation after
tomato, melon, watermelon, potato and onion.
Also, Tunisia is the third largest producer of pepper
after Nigeria and Egypt and the third largest
exporter (in terms of tonnage) after Morocco and
South Africa (Boughalleb and El Mahjoub, 2006).
Many pepper cultivars are cultivated in Tunisia , in
fields or under heated plastic tunnels, in costal and
southern regions. The present study evaluates the
effect of NaCl treatments on germination and early
growth parameters in three pepper cultivars which
are "Beldi", "Baklouti" and "Anaheim Chili" in order
to understand the reaction of young seedling to salt
stress and to select tolerant cultivar which can be
used in future experiences.
MATERIALS AND METHODS
Seed material
This study was conducted under controlled
environmental condition in the laboratory of the
Department of Agronomy and crop science of the
High Institute of Agriculture Chott Mariem , Sousse,
Tunisia. Seeds of three peppers cultivars ("Beldi",
"Baklouti" and "Anaheim Chili") were sterilized for
20 minutes in sodium hypochloride solution (5%),
then they were rinsed 3 times with distilled water
for 2 min. After sterilization under laminar flow, 20
seeds of each cultivar were transferred into sterile
Petri dishes (100 x100 mm dimensions) between
two layers of Watman filter paper and then
watered with 10 ml of distilled water (for control)
or 10 ml of a saline solution containing 2, 4, 6, 8, 10
and 12 g/l of NaCl and left to germinate at 25°C
(Figure 1.A). The Petri dishes were arranged in a
completely randomized block design with three
replications. Germinated seeds were recorded daily
during 14 days and the appearance of 2 mm or
more of radicle was considered as germination.
Parameters measured
Parameters measured in this study were:
Germination percentage (GP) which was
calculated based on the equation: GP = (Total
germinated seed) / (Total number of seed) (Ashraf
and Foolad, 2005).
Mean germination time (MGT) which was
calculated according to the equation of Ellis and
Roberts (1981) : MGT = Σ Dn / Σn. Where (n) is the
number of seeds which were germinated on day D,
and D is the number of days counted from the
beginning of germination.
Germination Index (GI) which was calculated as
described by the Association of Official Seed
Analysts (AOSA, 1983) as: GI= ∑ (Gt / Tt). GI=
[Number of germinated seeds in first count/ Day of
first count] +…+ [Number of germinated seeds in
final count/Day of final count).
The coefficient of velocity (CV) which was
calculated using the following formula:
CV= 100× [ΣNi / ΣNi Ti] where Ni is the number
of seeds germinated on i thdays, Ti is the number of
days from sowing (AOSA,1983). Plumule and radicle
length (cm) were measured with a graduated ruler.
Then, radicle and plumule fresh weight (FW) and
dry weight (DW) were determined with a precision
balance "Mettler". In order to determine dry weight
seedlings are separated and dried in an oven at
80°C for 48 hours.
Data analysis
Data were subjected to an analysis of variance,
using SPSS 13.0 software and the difference
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
16
Effect of Salt Stress (NaCl) on Germination...
between means were compared by Duncan
multiple range test at 5% level of probability.
Differences were considered significant at the 5%
level (means followed by different letters).
RESULTSTotal germination percentage
The results showed that different levels of
salinity have significant effect on pepper seed
germination. Many researchers have reported
similar results (Garcia et al., 1988 ; Ben Said, 2004).
In all of cultivars, there was a decrease in
germination percentage when the salt stress
increase. While, in this experiment, cultivars had
different response to the salinity (Figure 2). In
control, seed germination percentage varied
between 80% ("Baklouti" cultivar) and 90%
("Anaheim Chili" cultivar). At the highest salt
concentration (12 g/l), the maximum germination
percentage was observed in "Anaheim Chili"
cultivar (55%) and the lowest one in "Baklouti"
cultivar (5%). "Anaheim Chili" cultivar
demonstrated better tolerance to salt stress than
other cultivar for germination percentage.
Mean germination time
The results in Table 1 revealed that the
increasing salinity delayed significantly the mean
germination time (MGT) in all pepper cultivars. The
highest MGT was observed at 12 g/l of NaCl in
"Beldi" cultivar (9.25 days), and the lowest MGT
was observed at 0 g/l of NaCl in " Anaheim Chili"
cultivar (5.50 days). It has been reported that
salinity delays germination (Mensah and Ihenyen,
2009; Murillo-Amador et al., 2002; Rahman et al.,
2000). Similar result was also observed by
Redondo-Gomez et al. (2008), where MGT of
Gibraltar sea lavender (Limonium emarginatum L.)
was higher in salt stress condition.
Germination index
According to Table 1, salt stress (NaCl) had
significantly effect on germination index of all
pepper cultivars "Beldi", "Baklouti" and "Anaheim
Chili". The reduction gets stronger particularly at
the highest level of salt stress (12g/l). Similar results
were reported by Kaya et al. (2008) and Khan et al.
(2009) working on hot pepper. Among the three
pepper cultivar, the highest germination index
values were observed in "Anaheim Chili" cultivar in
all NaCl concentration.
Coefficient of Velocity
Considerable decrease was noted in Coefficient
of Velocity (Table 1), depending on NaCl
concentrations. In fact, coefficient of velocity varied
from 14,58 ("Baldi" cultivar) to 18,22 ("Anaheim
Chili" cultivar) in control. The addition of NaCl in
water reduced this coefficient .However, this
decrease was more pronounced for " Baldi " and
"Baklouti" cultivars than " Anaheim Chili" cultivar.
The minimum coefficient of velocity was found in
the high salinity treatment (12 g/l) and registered in
"Baldi" cultivar (10,83).
Radicle and plumule length
The effects of salinity stress on radicle and
plumule length of three pepper cultivars have been
showed in figure 3. Results showed a decrease in
radicle and plumule length with an increase in
salinity for the three pepper cultivars (Figure 1.B).
Plumule length was more suppressed than radicle
by salinity at all salt concentration levels. "Baldi"
cultivar seems to be the most sensitive cultivar and
"Anaheim Chili" the most tolerant one especially for
the highest concentrations of 10 and 12 g/l of NaCl.
Production of fresh and dry matter
Increased salt stress level also reduced the
production of fresh and dry matter in radicle and
plumule for all cultivars, although the reductions
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
17
Aloui et al.
were the lowest in "Anaheim Chili" cultivar (Figure
4 and 5). The most reduction (length and weight ) in
radicle and plumule was observed when salinity
reached 12 g/l of NaCl.
The highest dry weight of plumule and radicle
was observed for "Anaheim Chili" cultivar. While
"Baldi" cultivar showed highest reduction (22.75 %)
of plumule dry weight (from 0 to 12 g/l) and had
the highest reduction (50 %) of radicle dry weight.
Figure 1. (A) Germination in Petri dish of pepper seeds. (B) Effect of salt stress on radicle and plumule length of Pepper seedlings ("Anaheim Chili" cultivar).
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
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Effect of Salt Stress (NaCl) on Germination...
Figure 2. Effect of different salinity levels on Kinetics and total germination percentage of three
pepper cultivars: "Beldi", "Baklouti" and "Anaheim Chili"
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
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Aloui et al.
Table 1 : The effects of NaCl salinity on Mean Germination Time (MGT), Germination Index (GI) and Coefficient of Velocity (CV) of three paper cultivars : "Beldi" (BL) , "Baklouti" (BK) and "Anaheim Chili" (AC) .
NaCl (g l-1)
Cultivar 0 2 4 6 8 10
MGT
BL 6.93 e 7 de 7.47 cd 7.86 cd 8.38 bc 8.62 ab
BK 7.13 e 7.28 de 7.38 cd 7.57 cd 7.63 bc 8 ab
AC 5.50 e 6.50 de 6.68 cd 6.71cd 7.1bc 7.26 ab
GI
BL 2.45 ab 2.47 a 2.19 b 1.32 c 1.16 d 0.89 e
BK 2.85 ab 2.38 b 2.09 b 2.02 c 1.50 d 1.29 e
AC 3.16 ab 3.07 a 3.1b 2.58c 2.05d 1.59e
CV
BL 14.58a 14.28b 13.41bc 12.71cd 11.96cd 11.61de
BK 14.38a 13.73b 13.56bc 13.22cd 13.01cd 12.4de
AC 18.22a 15.58b 15.08bc 14.89cd 14.48cd 13.81de
Means followed by the same letters are not significantly different according to Duncan test at 5% level.
Figure 3. Effect of different level of salinity on radicle and plumule length of three pepper cultivars
"Beldi" (BL), "Baklouti" (BK) and "Anaheim Chili" (AC).
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
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Effect of Salt Stress (NaCl) on Germination...
Figure 4. Effect of different level of salinity on radicle fresh and dry weight (g) of three pepper
cultivars "Beldi" (BL), "Baklouti" (BK) and "Anaheim Chili" (AC).
Figure 5. Effect of different level of salinity on plumule fresh and dry weight (g) of three pepper
cultivars "Beldi" (BL), "Baklouti" (BK) and "Anaheim Chili" (AC).
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
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Aloui et al.
DISCUSSION
According to Ayaz et al. (2000), the reduction in
total seed germination under conditions of salt
stress is due to some metabolic disorders. It seems
that the decrease of total germination percentage is
related to the reduction in water absorption into
seeds (Hadas, 1977).
Gulzar and khan (2001) have reported that NaCl
stress prevents water absorption by seeds and
decreases significantly total germination
percentage. Keshavarzi (2011) and Keshavarzi et al.
(2011) have found the same result respectively on
some savory and spinach cultivars. In fact salinity
affects germination in two ways , first by decreasing
the osmotic potential which retard or prevent the
uptake of water and second by a toxic accumulation
of ions which damages the embryo. These results
have implications on the parameters of germination
such as total germination percentage, germination
index and the coefficient of velocity which are badly
affected. In addition, germination is delayed and
Mean Germination Time increased with salt stress.
The same results were reported by Zapata et al.
(2003) on lettuce (Lactuca sativa L.) and
Chartzoulokis and Klapaki (2000) on pepper.
Plumule was more suppressed than radicle by
salinity at all salt concentration levels. Similar
results were found by Keiffer and Ungar (1997),
Kaya et al. (2008) and Moud and Maghsoudo
(2008), who showed that shoots of seedlings were
more sensitive to salt stress than roots. This can be
the result of a fast osmotic adjustment occurring
in roots. The gradual decrease in plumule length
with increase in NaCl stress could be due to an
inhibitory effect of NaCl salt in shoot growth. It
seems that reduction in seedling fresh weight may
be due to a decrease in water uptake by seedling
under salinity. Similar results were reflected by
Anantharaju and Muthiah (2007).
Also, Jabeen et al. (2003) reported decline in
radicle growth under saline conditions. In addition,
Khan et al. (1997) indicated that the plumule length
significantly decreases with an increase in salt
concentrations. These results demonstrated that
the mechanism of tolerance to salinity is not yet
fully developed in these young seedlings of pepper.
and this characterize some sensitive plants in early
seedling stages (Almodares et al., 2007). Salinity has
both osmotic and specific ionic effects on seedlings
growth (Dioniso-Sese and Tobita, 2000). Similarly,
toxic ion accumulation (Na+ and Cl-) affect
negatively the metabolism of plants (Grieve and
Fujiyama, 1987). It has also been reported that
salinity suppresses the uptake of essential nutrients
like phosphorus (P) and potassium (K) (Nasim et al.,
2008), which could affect adversely seedlings
growth.
Similar results were reported by Munns (2002).
Vicente et al. (2004) demonstrated that the
reaction of plants to salt stress varied according to
the stage of plant development. That 's why these
pepper cultivar might be sensitive at early stage but
it could be tolerant at later stage growth. However,
based in these calculated parameters, the "Anaheim
Chili" is the most tolerant cultivar and can be used
in next selection programs.
CONCLUSION
In conclusion, this study demonstrated that seed
germination of pepper varied according to the
change in NaCl levels. The increase of salt stress
decreased all studied parameters on germination
and seedling growth and the increase of Mean
Germination Time (MGT). We found that "Anaheim
Chili" cultivar is the most resistant cultivar to salt
JOURNAL OF STRESS PHYSIOLOGY & BIOCHEMISTRY Vol. 10 No. 1 2014
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Effect of Salt Stress (NaCl) on Germination...
stress on germination stage. However, further
studies are needed to assess the agronomic
behavior of those cultivars during the later stages of
the culture.
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